• Title/Summary/Keyword: \beta-galactosidase

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The p53-p21Cip1/WAF1 Pathway Is Necessary for Cellular Senescence Induced by the Inhibition of Protein Kinase CKII in Human Colon Cancer Cells

  • Kang, Ji-Young;Kim, Jin Joo;Jang, Seok Young;Bae, Young-Seuk
    • Molecules and Cells
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    • v.28 no.5
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    • pp.489-494
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    • 2009
  • We have previously shown that the down-regulation of protein kinase CKII activity is tightly associated with cellular senescence of human fibroblast IMR-90 cells. Here, we examined the roles of p53 and $p21^{Cip1/WAF1}$ in senescence development induced by CKII inhibition using wild-type, isogenic p53-/- and isogenic p21-/- HCT116 human colon cancer cell lines. A senescent marker appeared after staining for senescence-associated ${\beta}$-galactosidase activity in wild-type HCT116 cells treated with CKII inhibitor or $CKII{\alpha}$ siRNA, but this response was almost abolished in p53- or $p21^{Cip1/WAF1}$-null cells. Increased cellular levels of p53 and $p21^{Cip1/WAF1}$ protein occurred with the inhibition of CKII. CKII inhibition upregulated p53 and $p21^{Cip1/WAF1}$ expression at post-transcriptional level and transcription level, respectively. RB phosphorylation significantly decreased in cells treated with CKII inhibitor. Taken together, this study shows that the activation of the $p53-p21^{Cip1/WAF1}$ pathway acts as a major mediator of cellular senescence induced by CKII inhibition.

Antimicrobial Characteristics of Paeonia lactiflora Pall. Extract Tested against Food-putrefactive Microorganisms (작약추출물의 식품변패미생물에 대한 항균특성)

  • Park, Ki-Duck;Cho, Sung-Hwan
    • Food Science and Preservation
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    • v.17 no.5
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    • pp.706-711
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    • 2010
  • Paeonia lactiflora Pall. was extracted with water and the Paeonia lactiflora Pall. extract (PLE) was tested for antimicrobial activities against Corynebacterium xerosis, Candida albicans, Listeria monocytogenes and Pseudomonas syringae. PLE showed pronounced antimicrobial effects at concentrations at or above 50 ${\mu}g$/mL. The activities were stable at $100^{\circ}C$ and over the pH range of 3-11. PLE may serve as a natural antimicrobial agent in food preservation. It is suggested that hydrophillic components in the extract synergistically perturb microbial membrane functions.

Influence of 10-Methacryloyloxydecyl Dihydrogen Phosphate on Cellular Senescence in Osteoblast-Like Cells

  • Ju Yeon Ban;Sang-Im Lee
    • Journal of dental hygiene science
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    • v.23 no.4
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    • pp.264-270
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    • 2023
  • Background: Resin-based dental materials release residual monomers or other substances from incomplete polymerization into the oral cavity, thereby causing adverse biological effects on oral tissue. 10-Methacryloyloxydecyl dihydrogen phosphate (10-MDP), an acidic monomer containing dihydrogen phosphate and methacrylate groups, is the most commonly used component of resin-based dental materials, such as restorative composite resins, dentin adhesives, and resin cements. Although previous studies have reported the cytotoxicity and biocompatibility in various cultured cells, the effects of resin monomers on cellular aging have not been reported to date. Therefore, this study aimed to investigate the effects of the resin monomer 10-MDP on cellular senescence and inflamm-aging in vitro. Methods: After stimulation with 10-MDP, MC3T3-E1 osteoblast-like cells were examined for cell viability by WST-8 assay and reactive oxygen species (ROS) production by flow cytometry. The protein and mRNA levels of molecular markers of aging were determined by western blotting and RT-PCR analysis, respectively. Results: Treatment with 0.05 to 1 mM 10-MDP for 24 hours reduced the survival of MC3T3-E1 cells in a concentration-dependent manner. The intracellular ROS levels in the 10-MDP-treated experimental group were significantly higher than those in the control group. 10-MDP at a concentration of 0.1 mM increased p53, p16, and p21 protein levels. Additionally, an aging pattern was observed with blue staining due to intracellular senescence-associated beta-galactosidase activity. Treatment with 10-MDP increased the levels of tumor necrosis factor-α, interleukin (IL)-1β, IL-6 and IL-8, however their expression was decreased by mitogen-activated-protein-kinase (MAPK) inhibitors. Conclusion: Taken together, these results suggest that the exposure of osteoblast-like cells to the dental resin monomer 10-MDP, increases the level of cellular senescence and the inflammatory response is mediated by the MAPK pathway.

Complete genome sequence of Enterococcus faecium strain AK_C_05 with potential characteristics applicable in livestock industry

  • Hyunok Doo;Jin Ho Cho;Minho Song;Eun Sol Kim;Sheena Kim;Gi Beom Keum;Jinok Kwak;Sriniwas Pandey;Sumin Ryu;Yejin Choi;Juyoun Kang;Hyeun Bum Kim;Ju-Hoon Lee
    • Journal of Animal Science and Technology
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    • v.66 no.2
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    • pp.438-441
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    • 2024
  • The Enterococcus faecium (E. faecium) strain AK_C_05 was isolated from cheonggukjang, the Korean traditional food, collected from a local market in South Korea. In this report, we presented the complete genome sequence of E. faecium strain AK_C_05. The genome of E. faecium strain AK_C_05 genome consisted of one circular chromosome (2,691,319 bp) with a guanine + cytosine (GC) content of 38.3% and one circular plasmid (177,732 bp) with a GC content of 35.48%. The Annotation results revealed 2,827 protein-coding sequences (CDSs), 18 rRNAs, and 68 tRNA genes. It possesses genes, which encodes enzymes such as alpha-galactosidase (EC 3.2.1.22), beta-glucosidase (EC 3.2.1.21) and alpha-L-arabinofuranosidase (EC 3.2.1.55) enabling efficient utilization of carbohydrates. Based on Clusters of Orthologous Groups analysis, E. faecium strain AK_C_05 showed specialization in carbohydrate transport and metabolism indicating the ability to generate energy using a variety of carbohydrates.

Phloroglucinol Enhances Anagen Signaling and Alleviates H2O2-Induced Oxidative Stress in Human Dermal Papilla Cells

  • Seokmuk Park;Ye Jin Lim;Hee Su Kim;Hee-Jae Shin;Ji-Seon Kim;Jae Nam Lee;Jae Ho Lee;Seunghee Bae
    • Journal of Microbiology and Biotechnology
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    • v.34 no.4
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    • pp.812-827
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    • 2024
  • Phloroglucinol (PG) is one of the abundant isomeric benzenetriols in brown algae. Due to its polyphenolic structure, PG exhibits various biological activities. However, the impact of PG on anagen signaling and oxidative stress in human dermal papilla cells (HDPCs) is unknown. In this study, we investigated the therapeutic potential of PG for improving hair loss. A non-cytotoxic concentration of PG increased anagen-inductive genes and transcriptional activities of β-Catenin. Since several anagen-inductive genes are regulated by β-Catenin, further experiments were performed to elucidate the molecular mechanism by which PG upregulates anagen signaling. Various biochemical analyses revealed that PG upregulated β-Catenin signaling without affecting the expression of Wnt. In particular, PG elevated the phosphorylation of protein kinase B (AKT), leading to an increase in the inhibitory phosphorylation of glycogen synthase kinase 3 beta (GSK3β) at serine 9. Treatment with the selective phosphoinositide 3-kinase/AKT inhibitor, LY294002, restored the increased AKT/GSK3β/β-Catenin signaling and anagen-inductive proteins induced by PG. Moreover, conditioned medium from PG-treated HDPCs promoted the proliferation and migration of human epidermal keratinocytes via the AKT signaling pathway. Subsequently, we assessed the antioxidant activities of PG. PG ameliorated the elevated oxidative stress markers and improved the decreased anagen signaling in hydrogen peroxide (H2O2)-induced HDPCs. The senescence-associated β-galactosidase staining assay also demonstrated that the antioxidant abilities of PG effectively mitigated H2O2-induced senescence. Overall, these results indicate that PG potentially enhances anagen signaling and improves oxidative stress-induced cellular damage in HDPCs. Therefore, PG can be employed as a novel therapeutic component to ameliorate hair loss symptoms.

The change on cell wall composition and physiological characteristic of astringent persimmon fruits by gamma irradiation (감마선 처리에 의한 떫은감 과육의 세포벽 성분 및 물성 변화)

  • Kim, Byung-Oh;Cha, Won-Seup;Ahn, Dong-Hyun;Cho, Young-Je
    • Food Science and Preservation
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    • v.22 no.4
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    • pp.512-519
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    • 2015
  • In this study, the effects of gamma-radiation treatment on cell wall composition and physiological characteristics of astringent persimmon fruit were investigated. The soluble tannin contents of gamma-radiated samples were reduced by the radiation treatment. The hardness of the radiated fruit was decreased when compared to non-radiated fruit. Alcohol-insoluble component of the cell wall in the radiated fruit was decreased from 39.3 mg/g to 37.2 mg/g. The water-soluble content of the radiated fruit was increased from 11.4 mg/g to 13.9 mg/g. The cell wall content of the non-radiated fruit was 26.6 mg/g whereas the cell wall content of radiated fruit was decreased to 23.1 mg/g. Due to the maturation of astringent persimmon fruit by gamma-radiation, water-soluble compounds were increased whereas decreasing in cell wall compounds. The contents of lignin, pectin, and cell wall were decreased from 0.82 mg/g and 3.56 mg/g to 0.77 mg/g and 3.14 mg/g, respectively. Acid-soluble hemicellulose content was decreased by gamma-radiation, while alkali-soluble hemicellulose and cellulose contents were increased. Activities of sotening enzyme as polygalacturonase, pectinesterase and $\beta$-galactosidase existed in persimmon fruit were increased by gamma-radiation. In the sensory evaluation, gamma-radition treated persimmon showed very low astringent taste when compared to the non-radiated fruit. In hardness test, the non-radiated persimmon maintained the hardness while gamma-treated persimmon showed softened outer layer due to the condensation of tannin during radiation treatment. Therefore, gamma-radiation treatment will be used for the removal of its astringency of persimmon fruit and for enhancement of its maturation.

Replicative Senescence of Periodontal Fibroblasts Induces the Changes in Gene Expression Pattern

  • Yi, Tac-Ghee;Jun, Ji-Hae;Min, Byung-Moo;Kim, Moon-Kyu;Kim, Gwan-Shik;Baek, Jeong-Hwa
    • International Journal of Oral Biology
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    • v.32 no.1
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    • pp.35-43
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    • 2007
  • Tooth loss in elderly is mainly caused by alveolar bone loss via severe periodontitis. Although the severity of periodontitis is known to be affected by age, the aging process or the genetic changes during the aging of periodontal tissue cells are not well characterized. In this study, we investigated the effect of in vitro aging on the change of gene expression pattern in periodontal fibroblasts. Gingival fibroblasts (GF) and periodontal ligament fibroblasts (PDL) were obtained from two young patients and replicative senescence was induced by sequential subcultivation. When more than 90% cells were positively stained with senescence-associated ${\beta},-galactosidase$, those cells were regarded as aged cells. In aged GF and PDL, the level of phosphorylated retinoblastoma (RB) and $p16^{INK4A}$ protein was significantly decreased and increased, respectively. However, the protein level of p53 and p21, well known senescence-inducing genes, did not increase in aged GF and PDL. Although $p27^{Kip1}$ and $p15^{INK4B}$, another cyclin-dependent kinase inhibitors, were reported to be involved in replicative senescence of human cells, they were decreased in aged GF and PDL. Because senescent cells showed flattened and enlarged cell shape and are known to have increased focal adhesion, we examined the protein level of several integrins. Aged GF and PDL showed increased protein level of integrin ${\alpha}2$, ${\alpha}v$, and ${\beta}1$. When the gene expression profiles of actively proliferating young cells and aged cells were compared by cDNA microarray of 3,063 genes and were confirmed by reverse transcription-polymerase chain reaction, 7 genes and 15 genes were significantly and commonly increased and decreased, respectively, in aged GF and PDL. Among them, included are the genes that were known to be involved in the regulation of cell cycle, gene transcription, or integrin signaling. The change of gene expression pattern in GF and PDL was minimally similar to that of oral keratinocyte. These results suggest that $p16^{INK4A}/RB$ might be involved in replicative senescence of periodontal fibroblasts and the change of gene expression profile during aging process is cell type specific.

Antimutagenic Mechanism of Water Extract from Rehmannia glutinosa Liboshitz on 4-nitroquinoline 1-oxide Induced Mutagenesis n E. coli B.r (대장균에서 4-nitroquinoline 1-oride의 변이원성에 대한 숙지황 물추출물의 항돌연변이 작용특성)

  • 안병용;한종현;최동성
    • KSBB Journal
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    • v.16 no.5
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    • pp.486-492
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    • 2001
  • The antimutagenic mechanism of the fraction III(RG III)separated from the water extract of Rehmannia glutionosa was investigated by Escherichia. coli GW and B/r strains. RG-III treatment did not affect the ${\beta}$-galactosidase activity E. coli GW-1060, 1106, 1107 and 1105. These results indicated that RG-III did not induce RecA protein amplification and did not also prevent the proteolytic cleavage of LexA. The bio-antimutagenicity and survival effect of RG-III on 4-nitroquinoline 1-oxide(4NQO), N-methyl-N-nitor-N\`-nitrosoguanidine(MNING) were investigate by E. coli B/r strains with have different pathway of DNA repai. RG-III slightly increased the survival of 4NQO-treated WP2, WP2s, WP67, CM561, CM611 cells, but the reactivation of survival cannot ve explained by the repair mode. RG-III caused the decrease of mutagenicity and lethality treated with MNNG in ZA159 despite of the increase in WP2, WP2s, WP67, CW561, CM611. Compared with bio-antimutagenic effects of RG-III on 4NQO, greatly increased antimutagenic effects of RG-III were observed with all the E. coli B/r strains tested, but less active in ZA159. These results suggest that RG-III was identified as a blocking agent for preventing the 4NQO induced mutagenesis, and may act as chl-products.

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An Identification of Enterobacter sp. Isolated from Contaminated Ginseng and Inhibition Effect of Ginseng Saponin on Its Growth (오염된 인삼으로부터 분리된 Enterobacter sp.의 동정 및 인삼사포닌의 균 생육억제효과)

  • 곽이성;이종태;여운형
    • Journal of Food Hygiene and Safety
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    • v.17 no.1
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    • pp.26-30
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    • 2002
  • A bacterium isolated from contaminated white ginseng was indentified by using API kit and electron microscope. The isolate was determined as rod shaped bacterium having 0.6-1.0 ${\mu}{\textrm}{m}$ in diameter and 1.2-3.0 ${\mu}{\textrm}{m}$ in length. It had motility by flagellum. The isolate had $\beta$-galactosidase, arginine dihydrolase and omithin decarboxylase. It used citrate as sole carbon source but not produced H$_2$S. It also fermented glucose, manitol, sorbitol, rhamnose, sucrose, melibiose, arabinose and amygdalin. The isolate was identified as Enterobacter sp by the above API kit analysis and electron microscopy observation. Ginseng saponin was added to culture of Enterobacter sp. in order to investigate saponin's influence on its growth. The strain was incubated at 38$^{\circ}C$ for 3 days after addition of 0.05, 0.5, 2.0 and 4.0% (w/v) of saponin, respectively and the growth rates were investigated. The relative bacterial growth rates showed 75.0, 37.5, 7.5 and 0.5%, respectively, when compared with 100% of saponin non-added group. These results suggest that the growth of Enterobacter sp. is inhibited by saponin with the concentration dependency.

Mini-transposon을 사용한 단백질의 세포내 분포 결정

  • 최의열
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.134-134
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    • 1993
  • Tn3 Transposon을 이용한 Shuttle Mutagenesis방법으로 효모의 Genome 상에 무작위적으로 $\beta$-gal 표지 유전자를 삽입하고 효모생활사의 각 세포시기 마다 특이하게 발현되는 유전자를 X-Gal plate상에서 찾아내고 이들 효모 유전자의 단백질이 세포내에 어떤 위치에 분포하는가를 간접 면역현미경법으로 추적해보았다. 먼저 효모의 genomic library를 38bp의 Tn3 Termial repeat를 가지고 있지 않은 pHSS6 Vector에 patial fill-in 방법으로 조성하였으며 최종적으로 20 Genome equivalent에 해당하는 18개 Pool의 genomic library를 만들었다. 이들 library를 조사하여 본 결과 모든 클론이 평균 3kb 크기의 insert를 가지고 있었으며 이는 99.99%의 효묘 genome을 대표하였다. 특정한 유전자의 발현을 알아보기 위해 먼저 mini-Tn3로 shuttle mutagenesis를 실시하고 vegetative growth동안 발현되는 유전자를 X-gal을 사용하여 골라내었다. 지금까지 16823개의 클론을 조사하였는데 이중 13%에 해당하는 2187개가 X-gal plate상에서 양성반응을 보여주었다. 양성반응을 보여주는 융합단백질의 세포내 분포틀 anti-$\beta$-galactosidase 항체를 사용하여 추적해보았다. 항체론 이용한 형광염색결과 약 70%의 세포가 background이상의 염색을 보여주었으며 이중 novel한 염색 pattern을 나타내는 클론도 다수 탐지되었다. 이상의 결과를 종합하면 Tn3틀 이용한 Shuttle Mutagenesis 방법으로 지금까지 전통적인 유전학적인 접근 방식으로 탐지되지 않았던 다수의 새로운 효모 유전자를 찾아낼 수 있을 것으로 사료된다.tamine제중 triprolidine이 $K_{M}$ /K$_{H}$ 비가 가장 높았고 diphenidol이 가장 낮았다. 이상의 결과로 보아 항 histamine제의 muscarinic receptor 차단작용은 이들 약물의 항 alleragy 효과에 필요한 작용이 아니며 본 실험에서 추정된 항 histamine제의 H$_1$-receptor와 muscarinic receptor에 대한 상대적 역가는 이들 약물의 선택과 평가에 중요한 지표가 될수 있을 것으로 생각된다.ing ischemic insults. The nature of the receptor is being explored by molecular genetic techniques, and we have recently cloned two of the major subunits; some of the data will be presented.LIFO, 우선 순위 방식등을 선택할 수 있도록 확장하였다. SIMPLE는 자료구조 및 프로그램이 공개되어 있으므로 프로그래머가 원하는 기능을 쉽게 추가할 수 있는 장점도 있다. 아울러 SMPLE에서 새로이 추가된 자료구조와 함수 및 설비제어 방식등을 활용하여 실제 중형급 시스템에 대한 시뮬레이션 구현과 시스템 분석의 예를 보인다._3$", chain segment, with the activation energy of carriers from the shallow trap with 0.4[eV], in he amorphous regions.의 증발산율은 우기의 기상자료를 이용하여 구한 결과 0.05 - 0.10 mm/hr 의 범위로서 이로 인한 강우손실량은 큰 의미가 없음을 알았다.재발이 나타난 3례의 환자를 제외한 9례 (75%)에서는 현재까지 재발소견을 보이지 않고 있다.

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